Journal: Journal of Oral Microbiology

Article Title: Aggregatibacter actinomycetemcomitans exacerbates colitis and perturbs the gut microbiota in a murine model​

doi: 10.1080/20002297.2026.2613536

Figure Lengend Snippet: Oral administration of A. actinomycetemcomitans was associated with aggravated intestinal barrier impairment and elevated levels of inflammatory mediators in mice with colitis. (A) Relative mRNA expression levels of pro-inflammatory cytokines in colon tissue were normalized to Gapdh (2 -ΔΔCt method). (B) Serum concentrations of IL-6, IL-1β, and TNF- α measured by ELISA. (C) Representative immunofluorescence images of colon sections stained for the tight junction protein ZO-1 (green) and the mucin MUC-2 (red). Nuclei were counterstained with DAPI (blue). Scale bars: 200 μm. Quantification of ZO-1 and MUC-2 signal intensity was performed on at least 9 non-overlapping fields per mouse using ImageJ software. Data are representative of two independent experiments. Data are shown as mean ± SEM. Statistical analysis was performed using two-sided one-way ANOVA followed by the least significant difference (LSD) post–hoc test or two-sided unpaired Student’s t-test and Dunnett T3 (uneven variance). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The ZO-1 and MUC-2 protein-targeting rabbit antibodies (Servicebio, China) were applied to the 4 μm-thick tissue sections, which were subsequently incubated overnight at 4 °C, washed with phosphate-buffered saline, and then treated with anti-rabbit fluorescent secondary antibodies (Servicebio, China) at 37 °C for 1 h. Following washing, the tissue nuclei were re-stained with 4-methyl-6-methyl-2-phenylindole (DAPI).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining, Software